Symcap
Teegan Innis
December 29, 2016
Project Summary
This project aimed to look at the spatial variability of color morph and Symbiodinium clades C and D present in the Kane’ohe Bay, O’ahu, Hawai’i population of Montipora capitata. We investigated the distributions at scales ranging from location within the bay to location on an individual reef. We also looked at differences among reef types (fringing vs. patch) and depths. Heterogeneous mixtures of symbiont clades were considered in the analysis for spatial patterns. By investigating spatial variability of Symbiodinium, we furthered the understanding of potential stress-response in Kane’ohe Bay.
Library Packages
knitr::opts_knit$set(root.dir = normalizePath(".."))
# setwd("~/Symcap") #This will not work on other people's computers and you should not need this - Rproj automatically sets wd to root of repository
# I don't thing all of these packages are actually being used in the code...
library(data.table)
library(lsmeans)
library(devtools)
library(plyr)
library(reshape2)
library(popbio)
library(RgoogleMaps)
library(plotrix)
library(zoo)
library(rgdal)
library(car)
library(scales)
library(png)
library(pixmap)
library(ecodist)
library(cluster)
library(fpc)
library(clustsig)
library(mapplots)
library(foreign)
library(nnet)
library(ggplot2)
library(mlogit)Import/Merge Collection and qPCR Data
# Import sample metadata
Coral_Data <- read.csv("Data/Collection Data/Coral_Collection.csv")
Coral_Data$Depth..m. <- as.numeric(as.character(Coral_Data$Depth..m.))
# Import qPCR data
source_url("https://raw.githubusercontent.com/jrcunning/steponeR/master/steponeR.R")
Mcap.plates <- list.files(path = "Data/qPCR_data", pattern = "txt$", full.names = T)
Mcap <- steponeR(files = Mcap.plates, delim="\t",
target.ratios=c("C.D"),
fluor.norm = list(C=2.26827, D=0),
copy.number=list(C=33, D=3))
Mcap <- Mcap$result
## Remove positive and negative controls from qPCR data
Mcap <- Mcap[grep("+", Mcap$Sample.Name, fixed=T, invert = T), ]
Mcap <- Mcap[grep("NTC", Mcap$Sample.Name, fixed = T, invert = T), ]
Mcap <- Mcap[grep("PCT", Mcap$Sample.Name, fixed = T, invert = T), ]
## Change "Sample.Name" column to "Colony"
colnames(Mcap)[which(colnames(Mcap)=="Sample.Name")] <- "Colony"
## Filter out detections without two technical replicates
Mcap$fail <- ifelse(Mcap$C.reps < 2 & Mcap$D.reps < 2, TRUE, FALSE)
fails <- Mcap[Mcap$fail==TRUE, ]
Mcap <- Mcap[which(Mcap$fail==FALSE),]
## Set C:D ratio to -Inf (when only D) and Inf (when only C)
Mcap$C.D[which(Mcap$C.reps<2)] <- -Inf
Mcap$C.D[which(Mcap$D.reps<2)] <- Inf
## Order qPCR data by Colony
Mcap <- Mcap[with(Mcap, order(Colony)), ]
## Calculate proportion C and proportion D
Mcap$propC <- Mcap$C.D / (Mcap$C.D+1)
Mcap$propD <- 1-Mcap$propC
Mcap$propD[which(Mcap$C.D==-Inf)] <-1
Mcap$propC[which(Mcap$C.D==-Inf)] <-0
Mcap$propD[which(Mcap$C.D==Inf)] <-0
Mcap$propC[which(Mcap$C.D==Inf)] <-1
## Categorize each sample as C-dominated or D-dominated
Mcap$Dom <- ifelse(Mcap$propC>Mcap$propD, "C", "D")
# Merge sample metadata with qPCR data
Symcap<-merge(Coral_Data, Mcap, by="Colony", all=T)
Symcap <- Symcap[with(Symcap, order(Colony)), ]
## Define category for symbiont mixture
Symcap$Mix <- factor(ifelse(Symcap$propC>Symcap$propD, ifelse(Symcap$propD!=0, "CD", "C"), ifelse(Symcap$propD>Symcap$propC, ifelse(Symcap$propC!=0, "DC", "D"), NA)), levels = c("C", "CD", "DC", "D"))
## Define reef.area category as either Top or Slope
Symcap$Reef.Area <- ifelse(Symcap$Reef.Area!="Top", yes = "Slope", no = "Top")
# Import color data from each observer
colscores <- read.csv("Data/Collection Data/Color_Scores.csv")
## Determine color scores by majority rule, and number of agreements
colscores$majority <- apply(colscores[,-1], 1, function(x) names(table(x))[which.max(table(x))])
colscores$n.agree <- apply(colscores[,-c(1,7)], 1, function(x) max(table(x)))
## Set which set of color scores will be used in analysis (=majority)
Symcap <- merge(Symcap, colscores[,c("Colony","majority","n.agree")], all=T)
Symcap$Color.Morph <- Symcap$majorityChange Bay Area for 18, 26 and F7-18
# PROVIDE RATIONALE FOR MAKING THESE CHANGES
Symcap$Bay.Area[which(Symcap$Reef.ID=="26")] <- "Central"
Symcap$Bay.Area[which(Symcap$Reef.ID=="18")] <- "Southern"
Symcap$Bay.Area[which(Symcap$Reef.ID=="F7-18")] <- "Southern"Adjust Depth by Mean Sea Level
To account for the influence of tides, depth was adjusted according to the differences in mean sea level from the recorded sea level on each collection day to the nearest 6-minute interval. Mean sea level was obtained from NOAA daily tide tables for Moku o Lo’e.
JuneTide=read.csv("Tide Tables/Station_1612480_tide_ht_20160601-20160630.csv")
JulyTide=read.csv("Tide Tables/Station_1612480_tide_ht_20160701-20160731.csv")
AugustTide=read.csv("Tide Tables/Station_1612480_tide_ht_20160801-20160812.csv")
Tide<-rbind(JuneTide, JulyTide, AugustTide)
Tide$Time <- as.POSIXct(Tide$TimeUTC, format="%Y-%m-%d %H:%M:%S", tz="UTC")
attributes(Tide$Time)$tzone <- "Pacific/Honolulu"
Symcap$Time2 <- as.POSIXct(paste(as.character(Symcap$Date), as.character(Symcap$Time)),
format="%m/%d/%y %H:%M", tz="Pacific/Honolulu")
Symcap$Time=Symcap$Time2
# Add estimated times for missing values
Symcap$Time[170] <- as.POSIXct("2016-06-14 12:07:00")
Symcap$Time[177] <- as.POSIXct("2016-06-14 12:20:00")
Symcap$Time[178] <- as.POSIXct("2016-06-14 12:22:00")
Symcap$Time[180] <- as.POSIXct("2016-06-14 13:08:00")
Symcap$Time[187] <- as.POSIXct("2016-06-14 12:42:00")
Symcap$Time[188] <- as.POSIXct("2016-06-14 12:44:00")
Symcap$Time[206] <- as.POSIXct("2016-06-16 13:10:00")
Symcap$Time[208] <- as.POSIXct("2016-06-16 13:24:00")
Symcap$Time[211] <- as.POSIXct("2016-06-16 12:37:00")
Symcap$Time[218] <- as.POSIXct("2016-06-16 12:27:00")
Symcap$Time[448] <- as.POSIXct("2016-07-16 13:32:00")
Round6 <- function (times) {
x <- as.POSIXlt(times)
mins <- x$min
mult <- mins %/% 6
remain <- mins %% 6
if(remain > 3L) {
mult <- mult + 1
} else {
x$min <- 6 * mult
}
x <- as.POSIXct(x)
x
}
Symcap$Time.r <- Round6(Symcap$Time)
Tide$Time.r <- Tide$Time
merged<-merge(Symcap, Tide, by="Time.r", all.x=T)
merged$newDepth <- merged$Depth..m.- merged$TideHTCoral Collection Map
Collection sites represented on this map indicate the 9 fringe and 16 patch reef sites for a total of 25 collection reefs. In total, 707 colonies were collected for the analysis.
# Define map area and get satellite data from Google using RgoogleMaps
KB <- c(21.46087401, -157.809907)
KBMap <- GetMap(center = KB, zoom = 13, maptype = "satellite", SCALE = 2, GRAYSCALE = FALSE)
# WHAT IS THE PURPOSE OF GENERATING MAP, SAVING TO FILE, THEN LOADING FROM FILE?
save(KBMap, file = "KBMap.Rdata")
load("KBMap.Rdata")
# Get representative coordinates for each reef based on sample GPS data
Latitude=aggregate(Latitude~Reef.ID, data=Symcap, FUN = mean)
Longitude=aggregate(Longitude~Reef.ID, data = Symcap, FUN=mean)
XY<-merge(Latitude, Longitude, by="Reef.ID", all=T)
newcoords <- LatLon2XY.centered(KBMap, XY$Latitude, XY$Longitude, zoom=13)
XY$X <- newcoords$newX
XY$Y <- newcoords$newY
XY <- subset(XY, Reef.ID!="37") # WHAT IS THIS DOING? WHY?
# Plot KBay Map
par(oma=c(3,3,0,0))
PlotOnStaticMap(KBMap, XY$Latitude, XY$Longitude, col=153, pch=21, bg="#FF7F50", lwd=2, cex = 1.2)
axis(1, at = LatLon2XY.centered(KBMap, NA, c(-157.85, -157.81, -157.77))$newX, tcl=0.5, line = 0.5, col = "ghostwhite", col.ticks = "black", lwd = 1, outer = TRUE, labels = c("157.85°W", "157.81°W", "157.77°W"), padj = -2.5, cex.axis = 0.75)
axis(2, at = LatLon2XY.centered(KBMap, c(21.42, 21.46, 21.50), NA)$newY, tcl=0.5, line = 0.5, col = "ghostwhite", col.ticks = "black", lwd = 1, outer = TRUE, labels = c("21.42°N", "21.46°N", "21.50°N"), padj = 0.5, hadj = 0.60, las = 1, cex.axis = 0.75)
par(new=T, mar=c(14,21,0,0))
HI <- readOGR("coast_n83.shp", "coast_n83")
HI <- spTransform(HI, CRS("+proj=longlat +datum=NAD83"))
plot(HI, xlim=c(-158.3, -157.6), ylim=c(21.35, 21.6), lwd=0.4, col="gray", bg="white")
rect(-157.87, 21.41, -157.75, 21.52)
box()
Symbiont Dominance and Color Morph Across Geographic Scales
Significant Differences
Dominant Symbiont per Reef Area
## I DON'T SEE ANYWHERE IN THE MANUSCRIPT THAT ANY EFFECT OF "REEF.AREA" (=TOP VS SLOPE) IS REPORTED OR DISCUSSED. IF NOT IN MANUSCRIPT, DELETE THIS ANALYSIS FROM MARKDOWN.
results <- table(Symcap$Dom, Symcap$Reef.Area)
chisq.test(results)##
## Pearson's Chi-squared test with Yates' continuity correction
##
## data: results
## X-squared = 136.26, df = 1, p-value < 2.2e-16prop.table(results, margin = 2)##
## Slope Top
## C 0.7767857 0.3294574
## D 0.2232143 0.6705426par(mar=c(4, 4, 2, 6))
barplot(prop.table(results, margin = 2), col = c("gray10", "gray100"), xlab = "Reef Area", ylab = "Proportion of colonies")
legend("topright", legend=c("C-dom.", "D-dom."), fill=c("gray10", "gray100"), inset = c(-.2, 0), xpd = NA)
Dominant Symbiont per Color Morph
results=table(Symcap$Dom, Symcap$Color.Morph)
chisq.test(results)##
## Pearson's Chi-squared test with Yates' continuity correction
##
## data: results
## X-squared = 263.61, df = 1, p-value < 2.2e-16prop.table(results, margin = 2)##
## Brown Orange
## C 0.92352941 0.32513661
## D 0.07647059 0.67486339par(mar=c(4, 4, 2, 6))
barplot(prop.table(results, margin = 2), col = c("gray10", "gray100"), xlab = "Color Morph", ylab = "Symbiont Proportion")
legend("topright", legend=c("C", "D"), fill=c("gray10", "gray100"), inset = c(-.2, 0), xpd = NA)
Symbiont Community Composition per Color Morph
results=table(Symcap$Mix, Symcap$Color.Morph)
chisq.test(results)##
## Pearson's Chi-squared test
##
## data: results
## X-squared = 266.44, df = 3, p-value < 2.2e-16prop.table(results, margin = 2)##
## Brown Orange
## C 0.794117647 0.286885246
## CD 0.129411765 0.038251366
## DC 0.073529412 0.653005464
## D 0.002941176 0.021857923par(mar=c(4, 4, 2, 6))
barplot(prop.table(results, margin = 2), col = c("gray10", "gray50", "gray85", "gray100"), xlab = "Color Morph", ylab = "Symbiont Community Composition")
legend("topright", legend=c("C", "CD", "DC", "D"), fill=c("gray10", "gray50", "gray85", "gray100"), inset = c(-.2, 0), xpd = NA)
Symbiont Community Composition per Reef Area
Symcap$Reef.Area <- ifelse(Symcap$Reef.Area!="Top", yes = "Slope", no = "Top")
results=table(Symcap$Mix, Symcap$Reef.Area)
chisq.test(results)##
## Pearson's Chi-squared test
##
## data: results
## X-squared = 138.97, df = 3, p-value < 2.2e-16prop.table(results, margin = 2)##
## Slope Top
## C 0.678571429 0.275193798
## CD 0.098214286 0.054263566
## DC 0.214285714 0.651162791
## D 0.008928571 0.019379845par(mar=c(4, 4, 2, 6))
barplot(prop.table(results, margin = 2), col = c("gray10", "gray50", "gray85", "gray100"), xlab = "Reef Area", ylab = "Symbiont Community Composition")
legend("topright", legend=c("C", "CD", "DC", "D"), fill=c("gray10", "gray50", "gray85", "gray100"), inset = c(-.2, 0), xpd = NA)
Symbiont Community Composition per Dominant Symbiont
results=table(Symcap$Mix, Symcap$Dom)
chisq.test(results)##
## Pearson's Chi-squared test
##
## data: results
## X-squared = 707, df = 3, p-value < 2.2e-16prop.table(results, margin = 2)##
## C D
## C 0.86635945 0.00000000
## CD 0.13364055 0.00000000
## DC 0.00000000 0.96703297
## D 0.00000000 0.03296703par(mar=c(4, 4, 2, 6))
barplot(prop.table(results, margin = 2), col = c("gray 10", "gray 85", "gray 40", "gray100"), xlab = "Dominant Symbiont", ylab = "Symbiont Mixture Proportion")
legend("topright", legend=c("C", "CD", "DC", "D"), fill=c("gray10", "gray85", "gray40", "gray100"), inset = c(-.2, 0), xpd = NA)
When looking at D-only colonies, the Ct values are on the higher end (35 or greater) indicating poor amplification or the potential for C amplification being pushed back in the cycle. This is supported by the fact that 5 of the 9 D-only colonies had C present in 1 qPCR replicate.
Proportion of D When Present in Mixture
propD <- merged$propD[which(merged$propD > 0 & merged$propD < 1)]
hist(propD, xlab = "Proportion of Clade D", ylab = "Number of Samples", main = "", col = "gray75")
Color Morph per Reef Area
Symcap$Reef.Area <- ifelse(Symcap$Reef.Area!="Top", yes = "Slope", no = "Top")
results=table(Symcap$Color.Morph, Symcap$Reef.Area)
chisq.test(results)##
## Pearson's Chi-squared test with Yates' continuity correction
##
## data: results
## X-squared = 57.683, df = 1, p-value = 3.08e-14prop.table(results, margin = 2)##
## Slope Top
## Brown 0.5902004 0.2906977
## Orange 0.4097996 0.7093023par(mar=c(4, 4, 2, 6))
barplot(prop.table(results, margin = 2), col = c("gray10", "gray100"), xlab = "Reef Area", ylab = "Color Morph Proportion")
legend("topright", legend=c("brown", "orange"), fill=c("gray10", "gray100"), inset = c(-.2, 0), xpd = NA)
Color Morph per Bay Area
results=table(merged$Color.Morph, merged$Bay.Area)
prop.table(results, margin = 2)##
## Central Northern Southern
## Brown 0.5288462 0.4358974 0.4769737
## Orange 0.4711538 0.5641026 0.5230263chisq.test(results)##
## Pearson's Chi-squared test
##
## data: results
## X-squared = 3.5162, df = 2, p-value = 0.1724Color Morph per Reef ID
results=table(Symcap$Color.Morph, Symcap$Reef.ID)
chisq.test(results)##
## Pearson's Chi-squared test
##
## data: results
## X-squared = 34.081, df = 24, p-value = 0.08324Latitude=aggregate(Latitude~Reef.ID, data=Symcap, FUN = mean)
Longitude=aggregate(Longitude~Reef.ID, data = Symcap, FUN=mean)
XY<-merge(Latitude, Longitude, by="Reef.ID", all=T)
propCol=table(Symcap$Color.Morph, Symcap$Reef.ID)
propCol=prop.table(propCol, margin = 2)
propCol <- as.data.frame.matrix(propCol)
props <- data.frame(t(propCol))
props$Reef.ID <- rownames(props)
XY<-merge(XY, props, by="Reef.ID", all=T)
newcoords <- LatLon2XY.centered(KBMap, XY$Latitude, XY$Longitude, zoom=13)
XY$X <- newcoords$newX
XY$Y <- newcoords$newY
PlotOnStaticMap(KBMap, XY$Latitude, XY$Longitude)
rownames(XY) <- XY$Reef.ID
XY <- XY[, -1]
XY <- na.omit(XY)
apply(XY, MARGIN=1, FUN=function(reef) {
floating.pie(xpos = reef["X"], ypos = reef["Y"],
x=c(reef["Orange"], reef["Brown"]), radius = 7, col = c("orange", "sienna"))
})
Mantel Test for Spatial Autocorrelation
Latitude=aggregate(Latitude~Reef.ID, data=Symcap, FUN = mean)
Longitude=aggregate(Longitude~Reef.ID, data = Symcap, FUN=mean)
XY<-merge(Latitude, Longitude, by="Reef.ID", all=T)
propCol=table(Symcap$Color.Morph, Symcap$Reef.ID)
propCol=prop.table(propCol, margin = 2)
propCol <- as.data.frame.matrix(propCol)
props <- data.frame(t(propCol))
props$Reef.ID <- rownames(props)
XY<-merge(XY, props, by="Reef.ID", all=T)
reef.dists <- dist(cbind(XY$Longitude, XY$Latitude))
col.dists <- dist(XY$Brown)
set.seed(12456)
mantel(col.dists~reef.dists)## mantelr pval1 pval2 pval3 llim.2.5% ulim.97.5%
## -0.04784488 0.66800000 0.33300000 0.59100000 -0.15930224 0.01286751Color Morph per Bay Area Adjusted for Depth
To discount the vertical spatial plane (depth), these results are adjusted to “assume” that all colonies were collected from the same depth, therefore considering simply the coordinate plane as the distributional influence on the resulting values.
merged$Color <- ifelse(merged$Color.Morph=="Orange", 1, 0)
dat <- aggregate(data.frame(prop=merged$Color), by=list(Reef.ID=merged$Reef.ID), FUN=mean, na.rm=T)
mod <- glm(Color ~ newDepth + Reef.ID, data=merged)
library(lsmeans)
lsm <- lsmeans(mod, specs="Reef.ID")
res <- merge(dat, data.frame(summary(lsm))[,c(1:2)], by="Reef.ID")
colnames(res) <- c("Reef.ID", "Raw", "O.Adj")
Latitude=aggregate(Latitude~Reef.ID, data=Symcap, FUN = mean)
Longitude=aggregate(Longitude~Reef.ID, data = Symcap, FUN=mean)
XY<-merge(Latitude, Longitude, by="Reef.ID", all=T)
propCol=table(Symcap$Color.Morph, Symcap$Reef.ID)
propCol=prop.table(propCol, margin = 2)
propCol <- as.data.frame.matrix(propCol)
props <- data.frame(t(propCol))
props$Reef.ID <- rownames(props)
XY<-merge(XY, props, by="Reef.ID", all=T)
newcoords <- LatLon2XY.centered(KBMap, XY$Latitude, XY$Longitude, zoom=13)
XY$X <- newcoords$newX
XY$Y <- newcoords$newY
PlotOnStaticMap(KBMap, XY$Latitude, XY$Longitude)
XY2 <- merge(XY, res, by = "Reef.ID")
XY2$B.Adj <- 1-XY2$O.Adj
PlotOnStaticMap(KBMap, XY$Latitude, XY$Longitude)
rownames(XY2) <- XY2$Reef.ID
XY2 <- XY2[, -1]
XY2 <- na.omit(XY2)
apply(XY2, MARGIN=1, FUN=function(reef) {
floating.pie(xpos = reef["X"], ypos = reef["Y"],
x=c(reef["O.Adj"], reef["B.Adj"]), radius = 7,
col = c("orange", "sienna"))
})
Mantel Test for Spatial Autocorrelation
reef.dists <- dist(cbind(XY$Longitude, XY$Latitude))
col.dists <- dist(XY2$O.Adj)
set.seed(12456)
mantel(col.dists~reef.dists)## mantelr pval1 pval2 pval3 llim.2.5% ulim.97.5%
## 0.02284001 0.33100000 0.67000000 0.76900000 -0.05447766 0.09818949Symbiont Community Composition per Bay Area
results=table(Symcap$Mix, Symcap$Bay.Area)
prop.table(results, margin = 2)##
## Central Northern Southern
## C 0.50961538 0.61538462 0.49174917
## CD 0.08173077 0.05641026 0.09900990
## DC 0.39423077 0.30256410 0.40594059
## D 0.01442308 0.02564103 0.00330033chisq.test(results)##
## Pearson's Chi-squared test
##
## data: results
## X-squared = 14.719, df = 6, p-value = 0.02256Non-Significant Differences
Dominant Symbiont per Reef Type
##
## Pearson's Chi-squared test with Yates' continuity correction
##
## data: results
## X-squared = 0.77593, df = 1, p-value = 0.3784Dominant Symbiont per Bay Area
results=table(Symcap$Dom, Symcap$Bay.Area)
chisq.test(results)##
## Pearson's Chi-squared test
##
## data: results
## X-squared = 3.8852, df = 2, p-value = 0.1433Dominant Symbiont per Reef ID
results=table(Symcap$Dom, Symcap$Reef.ID)
chisq.test(results)##
## Pearson's Chi-squared test
##
## data: results
## X-squared = 34.598, df = 24, p-value = 0.07459Latitude=aggregate(Latitude~Reef.ID, data=Symcap, FUN = mean)
Longitude=aggregate(Longitude~Reef.ID, data = Symcap, FUN=mean)
XY<-merge(Latitude, Longitude, by="Reef.ID", all=T)
propDom=table(Symcap$Dom, Symcap$Reef.ID)
propDom=prop.table(propDom, margin = 2)
propDom <- as.data.frame.matrix(propDom)
props <- data.frame(t(propDom))
props$Reef.ID <- rownames(props)
XY<-merge(XY, props, by="Reef.ID", all=T)
newcoords <- LatLon2XY.centered(KBMap, XY$Latitude, XY$Longitude, zoom=13)
XY$X <- newcoords$newX
XY$Y <- newcoords$newY
PlotOnStaticMap(KBMap, XY$Latitude, XY$Longitude)
rownames(XY) <- XY$Reef.ID
XY <- XY[, -1]
XY <- na.omit(XY)
apply(XY, MARGIN=1, FUN=function(reef) {
floating.pie(xpos = reef["X"], ypos = reef["Y"],
x=c(reef["C"], reef["D"]), radius = 7, col = c("blue", "red"))
})
Mantel Test for Spatial Autocorrelation
Latitude=aggregate(Latitude~Reef.ID, data=Symcap, FUN = mean)
Longitude=aggregate(Longitude~Reef.ID, data = Symcap, FUN=mean)
XY<-merge(Latitude, Longitude, by="Reef.ID", all=T)
propDom=table(Symcap$Dom, Symcap$Reef.ID)
propDom=prop.table(propDom, margin = 2)
propDom <- as.data.frame.matrix(propDom)
props <- data.frame(t(propDom))
props$Reef.ID <- rownames(props)
XY<-merge(XY, props, by="Reef.ID", all=T)
reef.dists <- dist(cbind(XY$Longitude, XY$Latitude))
dom.dists <- dist(XY$C)
set.seed(12456)
mantel(dom.dists~reef.dists)## mantelr pval1 pval2 pval3 llim.2.5% ulim.97.5%
## 0.15974663 0.04000000 0.96100000 0.04500000 0.08875833 0.24328773pval1 indicates that there is a positive correlation between distance and difference among reefs in terms of dominant symbiont clade. Reefs that are closer in proximity to each other are more similar in terms of symbiont dominance. A positive mantelr value indicates a positive correlation.
Clustering of Similar Reefs
Though insignificant differences resulted when investigating symbiont dominance across reefs, reefs closer in proximity appear more similar than reefs further apart. Clustering was tested here.
doms <- hclust(dom.dists)
plot(doms, labels = XY$Reef.ID)
a <- hclust(reef.dists*dom.dists)
plot(a, labels = XY$Reef.ID)
Dominant Symbiont Plots
Raw Values
Latitude=aggregate(Latitude~Reef.ID, data=Symcap, FUN = mean)
Longitude=aggregate(Longitude~Reef.ID, data = Symcap, FUN=mean)
XY<-merge(Latitude, Longitude, by="Reef.ID", all=T)
newcoords <- LatLon2XY.centered(KBMap, XY$Latitude, XY$Longitude, zoom=13)
XY$X <- newcoords$newX
XY$Y <- newcoords$newY
XY <- subset(XY, Reef.ID!="37")
merged$Dominant <- ifelse(merged$Dom=="C", 0, 1)
dat <- aggregate(data.frame(prop=merged$Dominant), by=list(Reef.ID=merged$Reef.ID), FUN=mean, na.rm=T) #prop D/reef
mod <- glm(Dominant ~ newDepth + Reef.ID, data=merged)
mod2 <- glm(Dominant ~ newDepth * Reef.ID, data=merged)
library(lsmeans)
lsm <- lsmeans(mod, specs="Reef.ID")
lsm2 <- lsmeans(mod2, specs="Reef.ID")
res <- merge(dat, data.frame(summary(lsm))[,c(1:2)], by="Reef.ID")
res <- merge(res, data.frame(summary(lsm2))[,c(1:2)], by="Reef.ID")
colnames(res) <- c("Reef.ID", "raw.c", "c.adj", "c.adj.int")
XY2 <- merge(XY, res, by = "Reef.ID")
XY2$d.adj <- 1-XY2$c.adj
XY2$d.adj.int <- 1-XY2$c.adj.int
XY2$raw.d <- 1-XY2$raw.c
PlotOnStaticMap(KBMap, XY$Latitude, XY$Longitude)
rownames(XY2) <- XY2$Reef.ID
XY2 <- XY2[, -1]
XY2 <- na.omit(XY2)
apply(XY2, MARGIN=1, FUN=function(reef) {
floating.pie(xpos = reef["X"], ypos = reef["Y"],
x=c(reef["raw.c"], reef["raw.d"]), radius = 7,
col = c("red", "blue"))
})
Depth Interaction Adjustment
merged$Dominant <- ifelse(merged$Dom=="C", 0, 1)
dat <- aggregate(data.frame(prop=merged$Dominant), by=list(Reef.ID=merged$Reef.ID), FUN=mean, na.rm=T) #prop D/reef
mod <- glm(Dominant ~ newDepth + Reef.ID, data=merged)
mod2 <- glm(Dominant ~ newDepth * Reef.ID, data=merged)
library(lsmeans)
lsm <- lsmeans(mod, specs="Reef.ID")
lsm2 <- lsmeans(mod2, specs="Reef.ID")
res <- merge(dat, data.frame(summary(lsm))[,c(1:2)], by="Reef.ID")
res <- merge(res, data.frame(summary(lsm2))[,c(1:2)], by="Reef.ID")
colnames(res) <- c("Reef.ID", "raw.c", "c.adj", "c.adj.int")
XY2 <- merge(XY, res, by = "Reef.ID")
XY2$d.adj <- 1-XY2$c.adj
XY2$d.adj.int <- 1-XY2$c.adj.int
XY2$raw.d <- 1-XY2$raw.c
PlotOnStaticMap(KBMap, XY$Latitude, XY$Longitude)
rownames(XY2) <- XY2$Reef.ID
XY2 <- XY2[, -1]
XY2 <- na.omit(XY2)
apply(XY2, MARGIN=1, FUN=function(reef) {
floating.pie(xpos = reef["X"], ypos = reef["Y"],
x=c(reef["c.adj.int"], reef["d.adj.int"]), radius = 7,
col = c("red", "blue"))
})
Color Morph and Dominant Symbiont Interaction Plots
A multinomial logistic regression was performed on the interaction of color morph and dominant symbiont to discount the influence of depth on spatial distribution.
merged$Dominant <- ifelse(merged$Dom=="C", 0, 1)
dat <- aggregate(data.frame(prop=merged$Dominant), by=list(Reef.ID=merged$Reef.ID),
FUN=mean, na.rm=T) #prop D/reef
mod <- glm(Dominant ~ newDepth + Reef.ID, data=merged)
mod2 <- glm(Dominant ~ newDepth * Reef.ID, data=merged)
library(lsmeans)
lsm <- lsmeans(mod, specs="Reef.ID")
lsm2 <- lsmeans(mod2, specs="Reef.ID")
res <- merge(dat, data.frame(summary(lsm))[,c(1:2)], by="Reef.ID")
res <- merge(res, data.frame(summary(lsm2))[,c(1:2)], by="Reef.ID")
colnames(res) <- c("Reef.ID", "raw.c", "c.adj", "c.adj.int")
XY2 <- merge(XY, res, by = "Reef.ID")
XY2$d.adj <- 1-XY2$c.adj
XY2$d.adj.int <- 1-XY2$c.adj.int
XY2$raw.d <- 1-XY2$raw.c
manteltable = table(merged$Dom, merged$Color.Morph, merged$Reef.ID)
nc <- aggregate(interaction(merged$Color.Morph, merged$Dom),
by=list(merged$Reef.ID), FUN=table)
nc <- data.frame(Reef.ID=as.character(nc[,1]), prop.table(nc[,2], margin=1))
XY3 <- merge(XY2, nc, by="Reef.ID", all=T)
merged$ColDom <- interaction(merged$Color.Morph, merged$Dom)
model <- multinom(ColDom~newDepth+Reef.ID, merged)
means <- lsmeans(model, specs = "Reef.ID")
means## Warning in sqrt(.qf.non0(object@V, x)): NaNs produced
## Warning in sqrt(.qf.non0(object@V, x)): NaNs produced
## Warning in sqrt(.qf.non0(object@V, x)): NaNs produced
## Warning in sqrt(.qf.non0(object@V, x)): NaNs produced
## Warning in sqrt(.qf.non0(object@V, x)): NaNs produced
## Warning in sqrt(.qf.non0(object@V, x)): NaNs produced
## Warning in sqrt(.qf.non0(object@V, x)): NaNs produced
## Warning in sqrt(.qf.non0(object@V, x)): NaNs produced
## Warning in sqrt(.qf.non0(object@V, x)): NaNs produced
## Warning in sqrt(.qf.non0(object@V, x)): NaNs produced
## Warning in sqrt(.qf.non0(object@V, x)): NaNs produced
## Warning in sqrt(.qf.non0(object@V, x)): NaNs produced
## Warning in sqrt(.qf.non0(object@V, x)): NaNs producedpp <- fitted(model)
newdat <- data.frame(Reef.ID = levels(merged$Reef.ID),
newDepth = mean(merged$newDepth, na.rm=T))
pred <- predict(model, newdata = newdat, "probs")
new <- data.frame(Reef.ID=as.character(newdat[,1]), pred)
XY4 <- merge(XY3, new, by="Reef.ID", all=T)
PlotOnStaticMap(KBMap, XY4$Latitude, XY4$Longitude)
XY4 <- merge(XY3, new, by="Reef.ID", all=T)
XY4[XY4 == 0.000] <- 0.0000000001
rownames(XY4) <- XY4$Reef.ID
XY4 <- XY4[, -1]
XY4 <- na.omit(XY4)
apply(XY4, MARGIN=1, FUN=function(reef) {
floating.pie(xpos = reef["X"], ypos = reef["Y"],
x=c(reef["Brown.C.x"], reef["Orange.C.x"], reef["Brown.D.x"],
reef["Orange.D.x"]), radius = 7,
col = c("turquoise4", "steelblue1", "tomato", "coral"))
})
legend("bottomleft", legend=c("Brown-C", "Orange-C", "Brown-D", "Orange-D"), fill = c("turquoise4", "steelblue1", "tomato", "coral"))
par(new=T, mar=c(15,22,0,0))
HI <- readOGR("coast_n83.shp", "coast_n83")
HI <- spTransform(HI, CRS("+proj=longlat +datum=NAD83"))
plot(HI, xlim=c(-158.3, -157.6), ylim=c(21.35, 21.6), lwd=0.4, col="gray", bg="white")
rect(-157.9, 21.41, -157.75, 21.53)
box()
Raw proportion values for Color Morph and Dominant Symbiont at each reef not discounting the effect of depth.
Mantel Test for Spatial Autocorrelation
reef.dists <- dist(cbind(XY4$Longitude, XY4$Latitude))
dom.dists <- bcdist(cbind(XY4$Brown.C.x, XY4$Orange.C.x, XY4$Brown.D.x, XY4$Orange.D.x))
set.seed(12456)
mantel(dom.dists~reef.dists)## mantelr pval1 pval2 pval3 llim.2.5% ulim.97.5%
## 0.1916069 0.0210000 0.9800000 0.0270000 0.1247632 0.2573410PlotOnStaticMap(KBMap, XY4$Latitude, XY4$Longitude)
XY4 <- merge(XY3, new, by="Reef.ID", all=T)
XY4[XY4 == 0.000] <- 0.0000000001
rownames(XY4) <- XY4$Reef.ID
XY4 <- XY4[, -1]
XY4 <- na.omit(XY4)
apply(XY4, MARGIN=1, FUN=function(reef) {
floating.pie(xpos = reef["X"], ypos = reef["Y"],
x=c(reef["Brown.C.y"], reef["Orange.C.y"], reef["Brown.D.y"],
reef["Orange.D.y"]), radius = 7,
col = c("turquoise4", "steelblue1", "tomato", "coral"))
})
legend("bottomleft", legend=c("Brown-C", "Orange-C", "Brown-D", "Orange-D"), fill = c("turquoise4", "steelblue1", "tomato", "coral"))
par(new=T, mar=c(15,22,0,0))
HI <- readOGR("coast_n83.shp", "coast_n83")
HI <- spTransform(HI, CRS("+proj=longlat +datum=NAD83"))
plot(HI, xlim=c(-158.3, -157.6), ylim=c(21.35, 21.6), lwd=0.4, col="gray", bg="white")
rect(-157.9, 21.41, -157.75, 21.53)
box()
Adjusted values of Dominant Symbiont and Color Morph adjusted for the influence of depth.
Mantel Test for Spatial Autocorrelation
reef.dists <- dist(cbind(XY4$Longitude, XY4$Latitude))
dom.dists <- bcdist(cbind(XY4$Brown.C.y, XY4$Orange.C.y, XY4$Brown.D.y, XY4$Orange.D.y))
set.seed(12456)
mantel(dom.dists~reef.dists)## mantelr pval1 pval2 pval3 llim.2.5% ulim.97.5%
## 0.11488056 0.08300000 0.91800000 0.16500000 0.04445094 0.19470011Color and Dominant Symbiont per Bay Area
When considering the effect of bay area on the interaction of color morph and dominant symbiont, the proportion of b colonies dominated by D increases as you move from the north to the south of the bay. The proportion increases almost 6x, yet the small number of colonies makes this non-compelling.
Type=table(merged$ColDom, merged$Bay.Area)
prop.table(Type, margin = 2)##
## Central Northern Southern
## Brown.C 0.48076923 0.43589744 0.42574257
## Orange.C 0.11057692 0.23589744 0.16501650
## Brown.D 0.04807692 0.00000000 0.05280528
## Orange.D 0.36057692 0.32820513 0.35643564chisq.test(Type)##
## Pearson's Chi-squared test
##
## data: Type
## X-squared = 20.668, df = 6, p-value = 0.002104Color Morph per Reef Type
##
## Pearson's Chi-squared test with Yates' continuity correction
##
## data: results
## X-squared = 0.08426, df = 1, p-value = 0.7716Symbiont Community Composition per Reef Type
##
## Pearson's Chi-squared test
##
## data: results
## X-squared = 2.8371, df = 3, p-value = 0.4174Symbiont Community Composition per Reef ID
##
## Pearson's Chi-squared test
##
## data: results
## X-squared = 87.127, df = 72, p-value = 0.1081Symbiont Dominance and Color Morph Across Depths
Dominant Symbiont by Depth
merged$Dominant <- ifelse(merged$Dom=="C", 0, 1)
Dom1 <- subset(merged, !is.na(newDepth) & !is.na(Dominant))
results=glm(Dominant~newDepth, family = "binomial", data = merged)
anova(results, test = "Chisq")## Analysis of Deviance Table
##
## Model: binomial, link: logit
##
## Response: Dominant
##
## Terms added sequentially (first to last)
##
##
## Df Deviance Resid. Df Resid. Dev Pr(>Chi)
## NULL 705 942.15
## newDepth 1 129.01 704 813.13 < 2.2e-16 ***
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1logi.hist.plot(Dom1$newDepth, Dom1$Dominant, boxp = FALSE, type = "hist", col="gray", xlabel = "Depth (m)", ylabel = "", ylabel2 = "")
mtext(side = 4, text = "Frequency", line = 3, cex=1)
mtext(side = 4, text = "C D", line = 2, cex = 0.75)
mtext(side = 2, text = "Probability of clade C Symbiont", line = 3, cex = 1)
merged$DepthInt <- cut(merged$newDepth, breaks = 0:13)
merged$Dominant2 <- ifelse(merged$Dom=="C", 1, 0)
results=table(merged$Dominant2, merged$DepthInt)
results##
## (0,1] (1,2] (2,3] (3,4] (4,5] (5,6] (6,7] (7,8] (8,9] (9,10] (10,11]
## 0 149 33 27 23 14 4 3 2 2 1 0
## 1 73 52 83 92 46 27 15 12 16 8 5
##
## (11,12] (12,13]
## 0 1 0
## 1 0 1props <- prop.table(results, margin = 2)
par(mar=c(4, 4, 2, 6), lwd = 0.25)
barplot(props[,1:11], col = c(alpha("red", 0.25), alpha("blue", 0.25)),
xlab = "", ylab = "",
space = 0, xaxs="i", yaxs="i", axisnames = FALSE)
par(lwd=1)
legend("topright", legend=c("C", "D"), fill=c(alpha("blue", 0.25), alpha("red", 0.25)), inset = c(-.2, 0), xpd = NA)
par(new = T)
par(mar=c(4.2, 4, 2, 6))
results=glm(Dominant~newDepth, family = "binomial", data = merged)
fitted <- predict(results, newdata = list(newDepth=seq(0,11,0.1)), type = "response")
plot(fitted~seq(0,11,0.1), xaxs="i", yaxs="i", xlim=c(0,11), ylim=c(0,1), type="l", lwd = 3, xlab="Depth (m)", ylab="Probability of D-Dominance")
Bars indicate relative frequency of occurence of clade C vs. D dominance at 1m depth intervals when pooling all samples collected. Overlaid line indicates probability of a colony being D-dominated across depths.
Mixture by Depth
merged$Mixture <- ifelse(!merged$Mix=="C" & !merged$Mix=="D", 1, 0)
merged$Mixture2 <- ifelse(!merged$Mix=="C" & !merged$Mix=="D", 0, 1)
results=glm(Mixture~newDepth, family = "binomial", data = merged)
anova(results, test = "Chisq")## Analysis of Deviance Table
##
## Model: binomial, link: logit
##
## Response: Mixture
##
## Terms added sequentially (first to last)
##
##
## Df Deviance Resid. Df Resid. Dev Pr(>Chi)
## NULL 705 973.27
## newDepth 1 93.723 704 879.55 < 2.2e-16 ***
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1results=table(merged$Mixture2, merged$DepthInt)
results##
## (0,1] (1,2] (2,3] (3,4] (4,5] (5,6] (6,7] (7,8] (8,9] (9,10] (10,11]
## 0 154 44 39 28 18 8 4 2 5 2 1
## 1 68 41 71 87 42 23 14 12 13 7 4
##
## (11,12] (12,13]
## 0 1 0
## 1 0 1props <- prop.table(results, margin = 2)
par(mar=c(4, 4, 2, 6), lwd = 0.25)
barplot(props[,1:11], col = c(alpha("red", 0.25), alpha("blue", 0.25)),
xlab = "", ylab = "",
space = 0, xaxs="i", yaxs="i", axisnames = FALSE)
par(lwd=1)
legend("topright", legend=c("Mix", "Non-Mix"), fill=c(alpha("blue", 0.25), alpha("red", 0.25)), inset = c(-.23, 0), xpd = NA)
par(new = T)
par(mar=c(4.2, 4, 2, 6))
results=glm(Mixture~newDepth, family = "binomial", data = merged)
fitted <- predict(results, newdata = list(newDepth=seq(0,11,0.1)), type = "response")
plot(fitted~seq(0,11,0.1), xaxs="i", yaxs="i", xlim=c(0,11), ylim=c(0,1), type="l", lwd = 3, xlab="Depth (m)", ylab="Mixture Proportion")
Bars indicate relative frequency of occurrence of Mixtures vs. Non-Mixtures at 1m depth intervals when pooling all samples collected. The overlaid line indicates the probability of a colony being a mixture across depths.
Color Morph by Depth
merged$Color <- ifelse(merged$Color.Morph=="Orange", 1, 0)
results=glm(Color~newDepth, family = "binomial", data = merged)
anova(results, test = "Chisq")## Analysis of Deviance Table
##
## Model: binomial, link: logit
##
## Response: Color
##
## Terms added sequentially (first to last)
##
##
## Df Deviance Resid. Df Resid. Dev Pr(>Chi)
## NULL 706 979.08
## newDepth 1 47.199 705 931.88 6.413e-12 ***
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1Color <- subset(merged, !is.na(newDepth) & !is.na(Color))
logi.hist.plot(independ = Color$newDepth, depend = Color$Color, type = "hist", boxp = FALSE, ylabel = "", col="gray", ylabel2 = "", xlabel = "Depth (m)")
mtext(side = 4, text = "Frequency", line = 3, cex=1)
mtext(side = 4, text = "Brown Orange", line = 2, cex = 0.75)
mtext(side = 2, text = "Probability of Orange Color Morph", line = 3, cex = 1)
merged$Color <- ifelse(merged$Color.Morph=="Orange", 0, 1)
results=table(merged$Color, merged$DepthInt)
results##
## (0,1] (1,2] (2,3] (3,4] (4,5] (5,6] (6,7] (7,8] (8,9] (9,10] (10,11]
## 0 153 37 51 58 24 11 9 1 6 1 1
## 1 69 48 59 57 36 20 9 13 13 8 4
##
## (11,12] (12,13]
## 0 1 0
## 1 0 1props <- prop.table(results, margin = 2)
par(mar=c(4, 4, 2, 6), lwd = 0.25)
barplot(props[,1:11], col = c(alpha("orange", 0.25), alpha("sienna", 0.25)),
xlab = "", ylab = "",
space = 0, xaxs="i", yaxs="i", axisnames = FALSE)
par(lwd=1)
legend("topright", legend=c("Brown", "Orange"), fill=c(alpha("sienna", 0.25), alpha("orange", 0.25)), inset = c(-.22, 0), xpd = NA)
par(new = T)
par(mar=c(4.2, 4, 2, 6))
merged$Color2 <- ifelse(merged$Color=="0", 1, 0)
results=glm(Color2~newDepth, family = "binomial", data = merged)
fitted <- predict(results, newdata = list(newDepth=seq(0,11,0.1)), type = "response")
plot(fitted~seq(0,11,0.1), xaxs="i", yaxs="i", xlim=c(0,11), ylim=c(0,1), type="l", lwd = 3, xlab="Depth (m)", ylab="Probability of Orange-Dominance")
Bars indicate relative frequency of b vs. o color morph dominance at 1m depth intervals when pooling all samples collected. The overlaid line indicates the probablity of a colony to be o across depths.
Color Morph and Dominant Symbiont Interaction
Dominant Symbiont per Depth and Bay Area
model=aov(Dominant~newDepth*Bay.Area, data = merged)
Anova(model, type = 2)## Anova Table (Type II tests)
##
## Response: Dominant
## Sum Sq Df F value Pr(>F)
## newDepth 25.026 1 125.1913 < 2e-16 ***
## Bay.Area 0.954 2 2.3870 0.09265 .
## newDepth:Bay.Area 1.555 2 3.8886 0.02092 *
## Residuals 139.933 700
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1Depth at which Orange switches from D to C Dominance
While b was always C-dominated, o was observed to switch from D to C dominance. The depth threshold where this switch occurred is represented here.
threshdepth <- function(color) {
df <- subset(merged, Color.Morph==color)
plot(df$Dominant2~df$newDepth, xlab="Depth (m)", ylab = "Probability of Clade C Symbiont",
main=color)
abline(h = 0.5, lty=2)
results=glm(Dominant2~newDepth, family = "binomial", data = df)
pval <- data.frame(coef(summary(results)))$`Pr...z..`[2]
mtext(side=3, text=pval)
newdata <- list(newDepth=seq(0,11,0.01))
fitted <- predict(results, newdata = newdata, type = "response")
lines(fitted ~ seq(0,11,0.01))
thresh <- ifelse(pval < 0.05,
newdata$newDepth[which(diff(sign(fitted - 0.5))!=0)], NA)
return(thresh)
}
sapply(levels(merged$Color.Morph), FUN=threshdepth)## list()merged$Dominant <- ifelse(merged$Dom=="C", 0, 1)
dat <- aggregate(data.frame(prop=merged$Dominant), by=list(Reef.ID=merged$Reef.ID),
FUN=mean, na.rm=T) #prop D/reef
mod <- glm(Dominant ~ newDepth + Reef.ID, data=merged)
mod2 <- glm(Dominant ~ newDepth * Reef.ID, data=merged)
library(lsmeans)
lsm <- lsmeans(mod, specs="Reef.ID")
lsm2 <- lsmeans(mod2, specs="Reef.ID")## NOTE: Results may be misleading due to involvement in interactionsres <- merge(dat, data.frame(summary(lsm))[,c(1:2)], by="Reef.ID")
res <- merge(res, data.frame(summary(lsm2))[,c(1:2)], by="Reef.ID")
colnames(res) <- c("Reef.ID", "raw.c", "c.adj", "c.adj.int")
XY2 <- merge(XY, res, by = "Reef.ID")
XY2$d.adj <- 1-XY2$c.adj
XY2$d.adj.int <- 1-XY2$c.adj.int
XY2$raw.d <- 1-XY2$raw.c
manteltable = table(merged$Dom, merged$Color.Morph, merged$Reef.ID)
nc <- aggregate(interaction(merged$Color.Morph, merged$Dom),
by=list(merged$Reef.ID), FUN=table)
nc <- data.frame(Reef.ID=as.character(nc[,1]), prop.table(nc[,2], margin=1))
XY3 <- merge(XY2, nc, by="Reef.ID", all=T)
merged$ColDom <- interaction(merged$Color.Morph, merged$Dom)
model <- multinom(ColDom~newDepth+Reef.ID, merged)## # weights: 108 (78 variable)
## initial value 978.723819
## iter 10 value 700.001242
## iter 20 value 684.814917
## iter 30 value 683.516711
## iter 40 value 683.175651
## iter 50 value 683.081045
## iter 60 value 683.060482
## iter 70 value 683.058377
## final value 683.058329
## convergedmeans <- lsmeans(model, specs = "Reef.ID")
means## Reef.ID prob SE df lower.CL upper.CL
## 10 0.25 4.032745e-10 78 0.25 0.25
## 13 0.25 NaN 78 NaN NaN
## 14 0.25 NaN 78 NaN NaN
## 18 0.25 NaN 78 NaN NaN
## 19 0.25 1.425791e-10 78 0.25 0.25
## 20 0.25 NaN 78 NaN NaN
## 21 0.25 NaN 78 NaN NaN
## 25 0.25 NaN 78 NaN NaN
## 26 0.25 1.840688e-10 78 0.25 0.25
## 30 0.25 NaN 78 NaN NaN
## 38 0.25 2.328306e-10 78 0.25 0.25
## 42 0.25 NaN 78 NaN NaN
## 46 0.25 NaN 78 NaN NaN
## 5 0.25 4.136431e-10 78 0.25 0.25
## Deep 0.25 NaN 78 NaN NaN
## F1-46 0.25 2.851581e-10 78 0.25 0.25
## F2-25 0.25 NaN 78 NaN NaN
## F3-18 0.25 2.328306e-10 78 0.25 0.25
## F4-34 0.25 2.328306e-10 78 0.25 0.25
## F5-34 0.25 3.292723e-10 78 0.25 0.25
## F6-Lilipuna 0.25 4.197414e-10 78 0.25 0.25
## F7-18 0.25 2.469542e-10 78 0.25 0.25
## F8-10 0.25 NaN 78 NaN NaN
## F9-5 0.25 1.164153e-10 78 0.25 0.25
## HIMB 0.25 NaN 78 NaN NaN
##
## Results are averaged over the levels of: ColDom
## Confidence level used: 0.95pp <- fitted(model)
depthmod <- multinom(ColDom ~ newDepth, merged)## # weights: 12 (6 variable)
## initial value 978.723819
## iter 10 value 745.177967
## final value 745.142241
## convergeddepthdat <- data.frame(newDepth = seq(0,12,0.5))
depthpred <- predict(depthmod, newdata=depthdat, "probs")
#plot(NA, xlim=c(0,12), ylim=c(0,1))
stackpoly(depthpred[,c(4,2,3,1)], stack=T, col=rev(c("burlywood4","burlywood3","darkorange3","darkorange")),
ylim=c(0,1), xaxlab=depthdat$newDepth, xlab="Depth", ylab="Probability")
legend("top", legend=c("Brown.C", "Brown.D", "Orange.C","Orange.D"), pch=22,
pt.bg=c("burlywood4","burlywood3","darkorange3","darkorange"), inset=-0.3, xpd=NA, bty="n")
df <- subset(merged, Color.Morph=="Orange")
results=glm(Dominant~newDepth, family = "binomial", data = df)
newdata <- list(newDepth=seq(0,11,0.01))
par(mar=c(4, 4, 2, 6))
fitted <- predict(results, newdata = newdata, type = "response")
plot(fitted ~ seq(0,11,0.01), ylim = c(0,1), type="l", col="orange", lwd=3, xlab="", ylab="", axisnames=FALSE)
abline(h = 0.5, lty=2)
df <- subset(merged, Color.Morph=="Brown")
results=glm(Dominant~newDepth, family = "binomial", data = df)
newdata <- list(newDepth=seq(0,11,0.01))
fitted <- predict(results, newdata = newdata, type = "response")
lines(fitted~seq(0,11,0.01), col="sienna", lwd=3)
mtext(side = 1, text = "Depth (m)", line = 3, cex = 1)
mtext(side = 2, text = "Probability of Clade D Symbiont", line = 3, cex = 1)
legend("topright", legend=c("Brown", "Orange"), fill=c("sienna", "orange"), inset = c(-.22, 0), xpd = NA)
Color Morph by Depth and Reef Type
model2=aov(Color~newDepth*Reef.Type, data = merged)
Anova(model2, type = 2)## Anova Table (Type II tests)
##
## Response: Color
## Sum Sq Df F value Pr(>F)
## newDepth 11.490 1 49.1052 5.693e-12 ***
## Reef.Type 0.188 1 0.8039 0.3702
## newDepth:Reef.Type 0.478 1 2.0448 0.1532
## Residuals 164.489 703
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1df <- subset(merged, Reef.Type=="Patch")
results=glm(Color2~newDepth, family = "binomial", data = df)
newdata <- list(newDepth=seq(0,11,0.01))
par(mar=c(4, 4, 2, 6))
fitted <- predict(results, newdata = newdata, type = "response")
plot(fitted ~ seq(0,11,0.01), ylim = c(0,1), type="l", col="dodgerblue3", lwd=3, xlab="", ylab="", axisnames=FALSE)
abline(h = 0.5, lty=2)
df <- subset(merged, Reef.Type=="Fringe")
results=glm(Color2~newDepth, family = "binomial", data = df)
newdata <- list(newDepth=seq(0,11,0.01))
fitted <- predict(results, newdata = newdata, type = "response")
lines(fitted~seq(0,11,0.01), col="brown1", lwd=3)
mtext(side = 1, text = "Depth (m)", line = 3, cex = 1)
mtext(side = 2, text = "Probability of Orange Color Morph", line = 3, cex = 1)
legend("topright", legend=c("Patch", "Fringe"), fill=c("dodgerblue3", "brown1"), inset = c(-.22, 0), xpd = NA)
Figures
Symbiont Community Composition per Dominant Symbiont
results=table(Symcap$Mix, Symcap$Dom)
chisq.test(results)
prop.table(results, margin = 2)
par(mar=c(4, 4, 2, 6))
barplot(prop.table(results, margin = 2), col = c(alpha("blue", 0.75), alpha("blue", 0.25), alpha("red", 0.25), alpha("red", 0.75)), xlab = "Dominant Symbiont", ylab = "Symbiont Mixture Proportion")
legend("topright", legend=c("C", "CD", "DC", "D"), fill=c(alpha("blue", 0.75), alpha("blue", 0.25), alpha("red", 0.25), alpha("red", 0.75)), inset = c(-.2, 0), xpd = NA)
When looking at D-only colonies, the Ct values are on the higher end of the spectrum (35 or greater) indicating poor amplification and the potential for C amplification being pushed back in the cycle. This is supported by the fact that 5 of the 9 D-only colonies had C present in 1 qPCR replicate.
Proportion of Clade D when Present
propD <- merged$propD[which(merged$propD > 0 & merged$propD < 1)]
propDHist <- subset(merged, propD > 0 & propD < 1)
propDHist$propD <- cut(propDHist$propD, breaks = 5)
DCol=table(propDHist$Color.Morph, propDHist$propD)
z <- with(subset(merged, propD==0), table(Color.Morph))
o <- with(subset(merged, propD==1), table(Color.Morph))
DCol <- cbind(z, DCol, o)
par(mar=c(4, 4, 2, 0))
bars <- barplot(DCol, xlab = "Clade D Proportion", ylab = "Number of Samples", main = "", col = c(alpha("sienna", 0.55), alpha("orange", 0.55)), axisnames = FALSE, space = c(0,0.3,0,0,0,0,0.3))
axis(side = 1, at = c(1.3, 2.3, 3.3, 4.3, 5.3, 6.3), labels = c(">0%", "20%", "40%", "60%", "80%", "<100%"))
axis(side = 1, at = c(0.5, 7.2), labels = c("All C", "All D"), tick = FALSE)
Depth Influence on Dominant Symbiont and Color Morph
par(mfrow=c(3,1))
merged$DepthInt <- cut(merged$newDepth, breaks = 0:13)
merged$Dominant <- ifelse(merged$Dom=="C", 0, 1)
merged$Dominant2 <- ifelse(merged$Dom=="C", 1, 0)
results=table(merged$Dominant2, merged$DepthInt)
results
props <- prop.table(results, margin = 2)
par(mar=c(2, 4, 2, 6), lwd = 0.25)
barplot(props[,1:11], col = c(alpha("red", 0.25), alpha("blue", 0.25)),
xlab = "", ylab = "",
space = 0, xaxs="i", yaxs="i", axisnames = FALSE)
par(lwd=1)
legend("topright", legend=c("C", "D"), fill=c(alpha("blue", 0.25), alpha("red", 0.25)), inset = c(0, 0), xpd = NA)
par(new = T)
par(mar=c(2.1, 4, 2, 6))
results=glm(Dominant~newDepth, family = "binomial", data = merged)
fitted <- predict(results, newdata = list(newDepth=seq(0,11,0.1)), type = "response")
plot(fitted~seq(0,11,0.1), xaxs="i", yaxs="i", xlim=c(0,11), ylim=c(0,1), type="l", lwd = 3, xlab="", ylab="Probability of D-Dominance")
merged$Color <- ifelse(merged$Color.Morph=="Orange", 0, 1)
results=table(merged$Color, merged$DepthInt)
results
props <- prop.table(results, margin = 2)
par(mar=c(3, 4, 1, 6), lwd = 0.25)
barplot(props[,1:11], col = c(alpha("orange", 0.25), alpha("sienna", 0.25)),
xlab = "", ylab = "Probability of Orange-Dominance",
space = 0, xaxs="i", yaxs="i", axisnames = FALSE)
par(lwd=1)
legend("topright", legend=c("Brown", "Orange"), fill=c(alpha("sienna", 0.25), alpha("orange", 0.25)), inset = c(0, 0), xpd = NA)
par(new = T)
par(mar=c(3.1, 4, 1, 6))
merged$Color2 <- ifelse(merged$Color=="0", 1, 0)
results=glm(Color2~newDepth, family = "binomial", data = merged)
fitted <- predict(results, newdata = list(newDepth=seq(0,11,0.1)), type = "response")
plot(fitted~seq(0,11,0.1), xaxs="i", yaxs="i", xlim=c(0,11), ylim=c(0,1), type="l", lwd = 3, xlab="", ylab="")
df <- subset(merged, Color.Morph=="Orange")
results=glm(Dominant~newDepth, family = "binomial", data = df)
newdata <- list(newDepth=seq(0,11,0.01))
par(mar=c(4, 4, 0, 6))
fitted <- predict(results, newdata = newdata, type = "response")
plot(fitted ~ seq(0,11,0.01), ylim = c(0,1), type="l", col="orange", lwd=3, xlab="Depth (m)", ylab="Probabilty of D-Dominance", axisnames=FALSE, xaxs = "i", yaxs = "i")
abline(h = 0.5, lty=2)
df <- subset(merged, Color.Morph=="Brown")
results=glm(Dominant~newDepth, family = "binomial", data = df)
newdata <- list(newDepth=seq(0,11,0.01))
fitted <- predict(results, newdata = newdata, type = "response")
lines(fitted~seq(0, 11, 0.01), col="sienna", lwd=3)
legend("topright", legend=c("Brown", "Orange"), fill=c("sienna", "orange"), inset = c(0, 0), xpd = NA)
#results=table(merged$Dominant2, merged$Color.Morph)
#chisq.test(results)
#prop.table(results, margin = 2)
#par(new=T, mar=c(10, 10, .5, 6.3))
#barplot(prop.table(results, margin = 2), col = c(alpha("red", 0.35), alpha("blue", 0.35)), xlab = "", ylab = "", yaxt = 'n')
#legend("topright", legend=c("C", "D"), fill=c(alpha("blue", 0.35), alpha("red", 0.35)), inset = c(0, 0), xpd = NA)Color Morph and Dominant Symbiont at each Reef
par(oma=c(3,3,0,0))
PlotOnStaticMap(KBMap, XY4$Latitude, XY4$Longitude)
axis(1, at = LatLon2XY.centered(KBMap, NA, c(-157.85, -157.81, -157.77))$newX, tcl=0.5, line = 0.5, col = "ghostwhite", col.ticks = "black", lwd = 1, outer = TRUE, labels = c("157.85°W", "157.81°W", "157.77°W"), padj = -2.5, cex.axis = 0.75)
axis(2, at = LatLon2XY.centered(KBMap, c(21.42, 21.46, 21.50), NA)$newY, tcl=0.5, line = 0.5, col = "ghostwhite", col.ticks = "black", lwd = 1, outer = TRUE, labels = c("21.42°N", "21.46°N", "21.50°N"), padj = 0.5, hadj = 0.60, las = 1, cex.axis = 0.75)
XY4 <- merge(XY3, new, by="Reef.ID", all=T)
XY4[XY4 == 0.000] <- 0.0000000001
rownames(XY4) <- XY4$Reef.ID
XY4 <- XY4[, -1]
XY4 <- na.omit(XY4)
apply(XY4, MARGIN=1, FUN=function(reef) {
floating.pie(xpos = reef["X"], ypos = reef["Y"],
x=c(reef["Brown.C.y"], reef["Orange.C.y"], reef["Brown.D.y"],
reef["Orange.D.y"]), radius = 7,
col = c("royalblue3", "cornflowerblue", "red", "rosybrown2"))
})
legend("bottomleft", legend=c("Brown-C", "Orange-C", "Brown-D", "Orange-D"), fill = c("royalblue3", "cornflowerblue", "red", "rosybrown2"))
par(new=T, mar=c(14,21,0,0))
HI <- readOGR("coast_n83.shp", "coast_n83")
HI <- spTransform(HI, CRS("+proj=longlat +datum=NAD83"))
plot(HI, xlim=c(-158.3, -157.6), ylim=c(21.35, 21.6), lwd=0.4, col="gray", bg="white")
rect(-157.87, 21.41, -157.75, 21.52)
box()
Adjusted values of Dominant Symbiont and Color Morph adjusted for the influence of depth.